Pharmaceutical preparation for the therapy of immune deficiency conditions

ABSTRACT

A pharmaceutical preparation for the therapy of immune deficiency conditions comprising an active principle--a peptide of the structure: H-L-Glu-L-Trp-OH and a pharmaceutically acceptable vehicle.

This application is a continuation of Ser. No. 08/337,341, filed Nov.10, 1994, now U.S. Pat. No. 5,538,951 , which is a continuation of08/194,189, filed Feb. 8, 1994, now abandoned, which is a continuationof Ser. No. 08/132,617, filed Oct. 7, 1993, now abandoned, which is acontinuation of Ser. No. 08/006,680, filed Jan. 21, 1993, now abandoned,which is a continuation of Ser. No. 07/856,802, filed Mar. 24, 1992, nowabandoned, which is a continuation of Ser. No. 07/415,283, filed Aug. 8,1989, now abandoned, which is a national stage application ofPCT/SU88/00255, filed Dec. 14, 1988, which has priority to U.S. Ser. No.4352833, filed Dec. 30, 1987.

FIELD OF THE INVENTION

The present invention relates to pharmacology and, in particular, to anovel pharmaceutical preparation for the therapy of immune deficiencyconditions.

STATE OF THE ART

Known in the art are preparations for the treatment of immune deficiencyconditions, in particular preparations of thymus obtained from animalraw materials: thymosin fraction 5 (cf. Goldstein A. L., Guha A., ZatzM. M., Hardy H. A., White A., Proc. Nat. Acad. Sci., USA, 1972, vol. 69,p. 1800-1803), thymalin (CH, A, 659586), T-activin (U.S. Pat. No.4,377,511). These preparations comprise a complex of substances of apolypeptide nature which are capable of controlling different stages ofproliferation and differentiation of T-lymphocytes. However, a practicaladministration of such preparations is substantially hampered due tocomplexity of processes for their manufacture, a low yield of activesubstances, and considerable variation of their physico-chemical andbiological properties. Furthermore, due to the presence of ballastcomponents in natural preparation of thymus, in some cases sidereactions appear in patients upon use of these preparations.

One of the most effective agents for the treatment of immune deficiencyconditions is thymalin comprising a complex of polypeptides with amolecular mass of 600-6,000 Dalton in a pharmaceutically acceptablevehicle such as glycine. It is administered in a dose of up to 10 mg ofan active principle once a day for 5 days. The content of activecomponents in this preparation causes its application in high doses (upto 50 mg per the treatment cause).

DISCLOSURE OF THE INVENTION

The present invention is directed to a pharmaceutical preparation forthe treatment of immune deficiency conditions incorporating such anactive principle of a peptide nature which would ensure a higherbiological activity of the preparation, its low toxicity and lack ofside effects.

This problem is solved by a pharmaceutical preparation or the therapy ofimmune deficiency conditions comprising an active principle of a peptidenature and a pharmaceutically acceptable vehicle. The preparationcontains, according to the present invention, as the active principle ofthe peptide nature, a peptide of the following structure:

    H-L-Glu-L-Trp-OH.

The pharmaceutical preparation according to the present invention forthe therapy of immune deficiency conditions has a high activity--in adose by 50-500 times lesser than that of thymalin it has a pronouncedimmunostimulant effect in the case of radiation induced immunedeficiency. Under the influence of the preparation there is observed aconsiderable stimulation of the production of lymphocytes andT-lymphocytes, as well as a clearly pronounced modulation of the ratiobetween subpopulations of immunocompetent cells. The LD₅₀ of thepreparation according to the present invention could not be established,since upon a 1000-times increase of the therapeutic dose (1 mg/kg), thelethal outcome was not revealed in any of the animals.

The preparation according to the present invention does not cause anyside effects.

It is advisable, that the pharmaceutical preparation according to thepresent invention would contain, when used as an injectable solution,the active principle in an amount of from 0.001 to 0.01% by weight.

In accordance with the present invention it is also advisable that thepharmaceutical preparation contain, as the pharmaceutically acceptablevehicle, a 0.9% aqueous solution of sodium chloride or a 0.5% solutionof novocain.

It is desirable, according to the present invention, that thepharmaceutical preparation in the form of tablets, suppositoria orcapsules contain the active principle in the amount of 0.1 mg pertablet, suppositorium or capsule.

It is also desirable, according to the present invention, that thepharmaceutical preparation in the form of tablets, suppositoria orcapsules contain the active principle in the amount of 0.1 mg pertablet, suppositorium or capsule.

It is also desirable, according to the present invention, that thepharmaceutical preparation in the form of tablets, suppositoria orcapsules would contain, as the pharmaceutically acceptable vehicle, afiller such as starch, glucose, glycine.

Other objects and advantages of the present invention will now becomemore fully apparent from the following detailed description of thepharmaceutical preparation for the therapy of immune deficiencyconditions.

BEST MODE FOR CARRYING OUT THE INVENTION

The pharmaceutical preparation for the therapy of immune deficiencyconditions according to the present invention incorporates a peptide ofthe following structure: H-L-Glu-L-Trp-OH and a pharmaceuticallyacceptable vehicle.

This peptide can be obtained by a conventional two-stage chemicalsynthesis in solution on the basis of an activated ether--protectedderivative of L-glutamic acid and L-tryptophane. After elimination ofthe protection groups and a chromatographic purification, the peptide islyophilized and obtained as an amorphous powder.

The peptide of the structure H-L-Glu-L-Trp-OH comprises a whitelyophilized powder readily soluble in water, dimethylformamide,insoluble in chloroform and ether. d!²² D=+12.6; C=0.5 H₂ O. R_(f) =0.65(butanol:acetic acid:water=3.1:1). The maximum of absorption in theUV-region is 275 ±5 nm. In the NMR spectrum at 500 MHz, 0.001 mol/l ofthe peptide solution, there are the following signals from thecorresponding aminoacid residues:

Trp-3.17; 3.37; 4.57; 7.16; 7.24; 7.71; 7.49

Glu-1.90; 1.96; 2.21; 3.72.

The above-given data clearly prove the structure of the synthesizedcompound--the active principle of the pharmaceutical preparationaccording to the present invention.

The active principle of the pharmaceutical preparation according to thepresent invention can be used as a free peptide and in the form ofwater-soluble salts thereof such as sodium, potassium, ammonium, zincsalts.

The preparation according to the present invention can be used indifferent pharmaceutical forms such as tablets, solutions(injection-intended and intranasal), drops, ointments and suppositoria.It is preferable to use the preparation according to the presentinvention as injectable solutions with a content of the active principleof from 0.001 to 0.01% by weight (0.0001-0.001 mg/kg of the body-mass,or 10-100 μg of the active principle per 1 ml of the solvent). As thepharmaceutically acceptable vehicle for the injection form, thepreparation can incorporate substantially any pharmaceuticallyacceptable solvent such as an aqueous solution of sodium chloride,distilled water, or a solution of novocain for injections, Ringer'ssolution, and a solution of glucose.

The preparation according to the present invention in the form oftablets and suppositoria preferably contains the active principle in theamount of 0.1 mg per one tablet or suppositorium. As thepharmaceutically acceptable vehicle for tablets, it is advisable to usestarch, glucose, glycine; for suppositoria, it is preferable to use, asthe pharmaceutically acceptable vehicle, any suitable pharmaceutic base.

The pharmaceutical preparation according to the present invention showsa high effectiveness in the treatment of immune deficiency conditions,it has a strong influence on the immunological responsiveness of theorganism, contributes to a considerable stimulation of the production oflymphocytes or T-lymphocytes and a pronounced modulation of the ratiobetween subpopulations of immunocompetent cells.

The preparation is not toxic, it is administered in low single andcourse doses and has a wide spectrum of the therapeutic action. Thepreparation according to the present invention can be widely employed inmedical practice for the therapy and prophylaxis of a whole number ofhuman diseases.

The pharmaceutical preparation according to the present invention forthe treatment of immune deficiency conditions has been experimentallystudied on animals and in clinics on human beings.

The biological activity of the preparation according to the presentinvention based on a dipeptide, as compared to the activity of thymalin,was studied in the cases of secondary immune deficiency conditions, inparticular those induced by radiation. The study was carried out on maleguinea pigs with a mass of 159-200 g. The animals were irradiated in aspecial unit with the dose of 1 Gr. One day after the irradiation, thetest groups of animals (10 guinea pigs in each group) were administeredwith thymalin and with the preparation according to the presentinvention in the doses of 0.0001-0.001 mg/kg in 0.5 ml of a 0.9% aqueoussolution of sodium chloride intramuscularly every day during 3 days. Thecontrol group of animals was administered with 0.5 ml of a 0.9% aqueoussolution of sodium chloride following the same scheme. On the 10th dayafter the irradiation in thymus and in the spleen, the number ofkaryocytes, T- and B-lymphocytes was determined by the method of rosetteformation.

The obtained results are shown in Tables 1 and 2 hereinbelow. It hasbeen found that the dipeptide in the dose of 0.001 mg/kg provides animmunostimulant effective by increasing, by more than 3 times, thenumber of karyocytes and T-lymphocytes in the thymus of the irradiatedanimals. In addition, the dipeptide in the dose of 0.001 mg/kgnormalizes the content of B-lymphocytes in the thymus and the spleen ofthe irradiated animals. Thymalin in all the studied doses does not exerta certain influence on the cell composition of the thymus and spleen ofthe irradiated animals.

                  TABLE 1                                                         ______________________________________                                        Comparative Study of the Effect of the Preparation According                  to the Present Invention and of Thymalin on the Cell                          Composition of Irradiated Guinea Pigs (X ± m)                                        Number of   Number of T-                                                                             Number of B-                                 Group of  Karyocytes, lymphocytes                                                                              lymphocytes                                  Animals   (thous./mg) (thous./mg)                                                                              (thous./mg)                                  ______________________________________                                        Intact    481.4 ± 21.3                                                                           378.5 ± 25.5                                                                          4.8 ± 0.3                                 Control    87.4 ± 13.2                                                                            49.6 ± 5.6                                                                           4.7 ± 0.6                                 (irradiation)                                                                 Dipeptide  89.3 ± 10.7                                                                            51.3 ± 6.7                                                                           5.3 ± 0.5                                 (0.00001 mg/kg)                                                               Dipeptide 139.8 ± 14.1.sup.x                                                                      89.4 ± 9.1.sup.x                                                                     4.9 ± 0.5.sup.x                           (0.00001 mg/kg)                                                               Dipeptide 275.8 ± 26.4.sup.x                                                                     194.7 ± 21.3.sup.x                                                                    3.3 ± 0.4.sup.x                           (0.001 mg/kg)                                                                 Thymalin   85.4 ± 11.6                                                                            53.6 ± 7.2                                                                           5.4 ± 0.6                                 (0.00001 mg/kg)                                                               Thymalin   90.4 ± 9.7                                                                             46.3 ± 5.7                                                                           5.3 ± 0.5                                 (0.0001 mg/kg)                                                                Thymalin   89.1 ± 13.6                                                                            47.9 ± 6.1                                                                           5.7 ± 0.7                                 (0.001 mg/kg)                                                                 ______________________________________                                         .sup.x statistically confident (P <0.05) as compared to the control.     

                  TABLE 2                                                         ______________________________________                                        Comparative Study of the Effect of the Preparation According                  to the Present Invention and of Thymalin on the Cell                          Composition of the Spleen of Irradiated Guinea Pigs (X ± m)                          Number of   Number of T-                                                                             Number of B-                                 Group of  Karyocytes, lymphocytes                                                                              lymphocytes                                  Animals   (thous./mg) (thous./mg)                                                                              (thous./mg)                                  ______________________________________                                        Intact    409.3 ± 25.6                                                                           40.4 ± 2.9                                                                            72.5 ± 5.4                                Control   147.9 ± 23.2                                                                           29.8 ± 3.4                                                                            51.4 ± 4.3                                (Irradiation)                                                                 Preparation of                                                                          150.6 ± 15.8                                                                           31.2 ± 3.6                                                                            50.9 ± 5.4                                the Invention                                                                 (0.00001 mg/kg)                                                               Preparation of                                                                          148.3 ± 16.0                                                                           30.4 ± 3.9                                                                            62.4 ± 6.1.sup.x                          the Invention                                                                 (0.0001 mg/kg)                                                                Preparation of                                                                          146.6 ± 18.1                                                                           29.9 ± 3.3                                                                            73.2 ± 5.1.sup.x                          the Invention                                                                 (0.001 mg/kg)                                                                 Thymalin  151.4 ± 16.7                                                                           32.0 ± 4.1                                                                            53.7 ± 6.3                                (0.00001 mg/kg)                                                               Thymalin  147.3 ± 15.1                                                                           31.7 ± 5.2                                                                            49.7 ± 5.8                                (0.0001 mg/kg)                                                                Thymalin  150.0 ± 16.6                                                                           28.9 ± 4.1                                                                            54.7 ± 5.2                                (0.001 mg/kg)                                                                 ______________________________________                                         .sup.x Statistically confident (P <0.05) as compared to the control.     

From the results obtained in an additional series of experiments undersimilar conditions, it has been found that thymalin provides animmunostimulating effect only in the dose of 0.1 mg/kg of the animal'sbody mass (see the data of Table 3 hereinbelow).

                  TABLE 3                                                         ______________________________________                                        Effect of Thymalin on Cell Composition of the Thymus and Spleen of            Irradiated Guinea Pigs (X ± m)                                             ______________________________________                                                    Number of Karyocytes (thous./mg)                                  Group of Animals                                                                            Thymus       Spleen                                             1             2            3                                                  ______________________________________                                        Intact        481.4 ± 21.3                                                                            409.3 ± 25.6                                    Control (irradiation)                                                                        87.4 ± 13.2                                                                            147.9 ± 23.2                                    Thymalin       89.3 ± 11.6                                                                            153.6 ± 17.1                                    (0.01 mg/kg)                                                                  Thymalin      101.4 ± 15.3                                                                            149.4 ± 16.0                                    (0.05 mg/kg)                                                                  Thymalin      280.7 ± 27.3.sup.x                                                                      154.3 ± 16.1                                    (0.1 mg/kg)                                                                   ______________________________________                                                  Number of Karyocytes                                                                          Number of Karyocytes                                Group of  (thous./mg)     (thous./mg)                                         Animals   Thymus     Spleen   Thymus Spleen                                   1         4          5        6      7                                        ______________________________________                                        Intact    378.5 ± 25.5                                                                          40.4 ± 2.9                                                                          4.8 ± 0.3                                                                         72.5 ± 5.4                            Control    49.6 ± 5.6                                                                           29.8 ± 3.4                                                                          5.7 ± 0.6                                                                         51.4 ± 4.3                            (irradiation)                                                                 Thymalin   51.3 ± 7.8                                                                           33.6 ± 5.7                                                                          5.9 ± 0.7                                                                         54.5 ± 6.3                            (0.01 mg/kg)                                                                  Thymalin   73.7 ± 14.1                                                                          26.4 ± 4.1                                                                          5.1 ± 0.6                                                                         61.7 ± 9.0                            (0.05 mg/kg)                                                                  Thymalin  206.7 ± 19.8.sup.x                                                                    30.1 ± 3.3                                                                          3.9 ± 0.5.sup.x                                                                   75.6 ± 6.2.sup.x                      (0.1 mg/kg)                                                                   ______________________________________                                         .sup.x Statistically confident as compared to the control (P <0.05).     

Therefore, the preparation according to the present invention in a doseby 50-500 times smaller than thymalin exhibits a clearly pronouncedimmunostimulant effect in the case of radiation induced immunedeficiency.

A comparative assessment of the effect produced by the preparationaccording to the present invention and that of thymalin on expression ofreceptors of thymocytes obtained from intact guinea pigs was made. It isknown that after treatment of lymphocytes with trypsin there occurs thedecomposition of the major portion of E-receptors on the surface ofcells, wherefore the percentage of T-lymphocytes is reduced which isrevealed in the reaction of rosette-formation (E-RFC). The addition ofthymus preparations to the trypsin-treated lymphocytes in therestoration of destroyed E-receptors which is revealed in an increasedpercentage of E-RFC (T-lymphocytes).

The preparation according to the present invention and thymalin wereadded to thymocytes obtained from 10 intact guinea pigs in a doseranging from 0.0001-1 μg/ml. The results of the study are shown in Table4 hereinbelow.

It has been found that the preparation according to the presentinvention and thymalin restored E-receptors of thymocytes treated withtrypsin. The preparation according to the present invention waseffective in a dose of from 0.01 to 1 μg/ml.

The assessment of clinico-immunological effectiveness of thepharmaceutical preparation according to the present invention was madeon 300 patients with purulent-inflammatory diseases of bones and softtissues (chronic post-traumatic osteomyelitis of long tubular bones,purulent-inflammatory complications of trauma and operativeinterventions in the case of diseases of the locomotor system, acute andchronic purulent-inflammatory diseases of bones and soft tissues ofmaxillofacial area, staphylococcal pyoderma).

                  TABLE 4                                                         ______________________________________                                        Effect of the Preparation According to the Present Invention                  on Expression of E-receptors of Thymocytes                                    (X ± m)                                                                    Preparation     EA - RFC (%)                                                                             P                                                  ______________________________________                                        Control         36.1 ± 3.1                                                                            --                                                 Preparation of the                                                                            37.4 ± 4.0                                                                            >0.05                                              Present Invention                                                             (0.0001 μg/ml)                                                             Preparation of the                                                                            41.4 ± 5.1                                                                            >0.05                                              Present Invention                                                             (0.001 μg/ml)                                                              Preparation of the                                                                            61.4 ± 5.3                                                                            <0.05                                              Present Invention                                                             (0.01 μg/ml)                                                               Preparation of the                                                                            62.7 ± 6.1                                                                            <0.05                                              Present Invention                                                             (1 μg/ml)                                                                  Thymalin        35.3 ± 3.9                                                                            >0.05                                              (0.0001 μg/ml)                                                             Thymalin        39.6 ± 4.1                                                                            >0.05                                              (0.001 μg/ml)                                                              Thymalin        38.7 ± 4.0                                                                            >0.05                                              (0.01 μg/ml)                                                               Thymalin        57.0 ± 4.7                                                                            <0.05                                              (1 μg/ml)                                                                  ______________________________________                                    

The pharmaceutical preparation according to the present invention wasintramuscularly or intranasally administered in a single dose of 100 μgfor 3-10 days (500 μg of the preparation on the average per course). Thecontrol group was composed of 200 patients with a similar pathology.

In the patients with purulent-inflammatory diseases of bones and softtissues of different localization, there has been revealed, to adifferent extent, of quantitative and especially functionalcharacteristics of the cell immunity and of non-specific resistance.Also noted is the violation of differentiation of subpopulations ofT-cells: a reduced number of T-helpers (OKT4⁺) and T-suppressors(OKT8⁺).

The use of the pharmaceutical preparation according to the presentinvention contributed to an improvement of the clinical progress of thedisease or to healing in 75.3-91.7% of patients which was revealed in areduced period of cleansing and healing of wounds, diminution of thearea of destruction of the bone tissue, in a faster arresting ofpurulent-inflammatory diseases of the skin, shortening of the treatmenttime by 20-30% as compared to the control patients. The clinicaleffectiveness of the preparation according to the present invention wasaccompanied by restoration of both quantitative and functionalcharacteristics of the T-system of immunity and non-specific resistance.

In none of the cases of clinical applications of the preparationaccording to the present invention toxic or allergic reactions werenoticed.

A comparative assessment of the effect of the preparation according tothe present invention and of thymalin was made on subpopulation oflymphocytes in patients with different pathologies, namely: forT-helpers (OKT4⁺), T-suppressors (OKT8⁺), coefficient of differentiationOKT4⁺ /OKT8⁺ and B-lymphocytes (Ig⁺) in patients with thymomegalia,thymectomy, rheumatoid arthritis, furunculosis and breast cancer (afterradiation therapy). In cultures of lymphocytes of the peripheral bloodof the patients, the percentage of subpopulations of lymphocytes wasdetermined by the immunofluorescent method using monoclonal antibodiesprior to and after introduction of thymalin and the preparation of thepresent invention in the most effective doses of 1 μg/ml and 0.01 μg/mlrespectively.

The results of the studies are shown in Tables 5 and 6.

                                      TABLE 5                                     __________________________________________________________________________    Effect of Thymalin on Subpopulations of Lymphocytes in Patients with          Different Pathologies (in the Numerator - the Starting Amount, in the         Denominator - After the Addition of Thymalin)                                              Number of Lymphocytes (%) X ± m                               Pathology                                                                            Number of Experiments                                                               T-Helpers (OKT4.sup.+)                                                               T-Suppressors (OKT8.sup.+)                                                            ##STR1##                                                                             B-Lymphocytes (Ig.sup.+)                   __________________________________________________________________________    Healthy                                                                              18                                                                                   ##STR2##                                                                             ##STR3##                                                                             ##STR4##                                                                              ##STR5##                                  Thymomegalia                                                                          8                                                                                   ##STR6##                                                                             ##STR7##                                                                             ##STR8##                                                                              ##STR9##                                  Thymectomy                                                                            4                                                                                   ##STR10##                                                                            ##STR11##                                                                            ##STR12##                                                                             ##STR13##                                 Rheumatoid Arthritis                                                                  4                                                                                   ##STR14##                                                                            ##STR15##                                                                            ##STR16##                                                                             ##STR17##                                 Furunculosis                                                                          6                                                                                   ##STR18##                                                                            ##STR19##                                                                            ##STR20##                                                                             ##STR21##                                 Breast Cancer (After Radiation Therapy)                                              10                                                                                   ##STR22##                                                                            ##STR23##                                                                            ##STR24##                                                                             ##STR25##                                  .sup.x Statistically confident (P < 0.05) as compared to the intial           characteristics.                                                         

                                      TABLE 6                                     __________________________________________________________________________    Effect of the Preparation According to the Present Invention on               Subpopulations of Lymphocytes in Patients with Different Pathologies                       Number of Lymphocytes (%) X ± m                               Pathology                                                                            Number of Experiments                                                               T-Helpers (OKT4.sup.+)                                                               T-Suppressors (OKT8.sup.+)                                                            ##STR26##                                                                            B-Lymphocytes (Ig.sup.+)                   __________________________________________________________________________    Healthy                                                                              18                                                                                   ##STR27##                                                                            ##STR28##                                                                            ##STR29##                                                                             ##STR30##                                 Thymomegalia                                                                          8                                                                                   ##STR31##                                                                            ##STR32##                                                                            ##STR33##                                                                             ##STR34##                                 Thymectomy                                                                            4                                                                                   ##STR35##                                                                            ##STR36##                                                                            ##STR37##                                                                             ##STR38##                                 Rheumatoid Arthritis                                                                  4                                                                                   ##STR39##                                                                            ##STR40##                                                                            ##STR41##                                                                             ##STR42##                                 Furunculosis                                                                          6                                                                                   ##STR43##                                                                            ##STR44##                                                                            ##STR45##                                                                             ##STR46##                                 Breast Cancer (After Radiation Therapy)                                              10                                                                                   ##STR47##                                                                            ##STR48##                                                                            ##STR49##                                                                             ##STR50##                                  .sup.x Statistically confident (P < 0.05) as compared to the starting         characteristic.                                                          

It has been shown that thymalin and the preparation according to thepresent invention (in a dose nearly 100 times as low as that ofthymalin) normalizes the ratio between subpopulations of lymphocytes inall groups of the examined patients.

Therefore, the results of the experimental study of the novelpharmaceutical preparation have shown its high efficiency in thetreatment of immune deficiency conditions. As compared to thymalin, thepreparation of this invention has a stronger (by about 50-500 times)effect on the immunological reactivity of the organism. The preparationgives rise to a considerable stimulation of the production oflymphocytes and T-lymphocytes, as well as to a clearly pronouncedmodulation of the ratio between subpopulations of immunocompetent cells.

The preparation according to the present invention can be used invarious pharmaceutical forms such as injectable solutions, tablets,suppositoria, ointments. In the case of using the preparation accordingto the present invention as injectable solutions, the latter contain theactive principle in an amount of from 0.001 to 0.01% by weight(0.0001-0.001 mg/kg of the body mass or 10-100 μg of the activeprinciple in 1 ml of the solvent). As the diluent the preparationaccording to the present invention preferably incorporates a 0.9%aqueous solution of sodium chloride, distilled water, a solution ofnovocain for injections, Ringer's solution, a solution of glucose.

The pharmaceutical preparation according to the present invention in theform of tablets and suppositoria preferably contains the activeprinciple in the amount of 0.1 mg per tablet or suppositorium. Fortablets and suppositoria as a pharmaceutical filler, use is made of anypharmaceutically acceptable vehicle. The preparation in the form of aninjectable solution is administered intramuscularly at the rate of oneinjection during 5 days. The intranasal administration of thepreparation is also effected during 5 days at the rate of one ampule orsyringe-tube.

The preparation according to the present invention does not have anyside effect and has no contraindications against its application. Thepharmaceutical forms of the preparation according to the presentinvention are prepared by conventional methods.

Industrial Applicability

The preparation according to the present invention is useful in themedical practice for the treatment and prevention of immune deficiencyconditions of the human organism.

We claim:
 1. A method for increasing the number of T-lymphocytes,B-lymphocytes or karyocytes in a non-human animal in need thereofcomprising administering to the animal a pharmaceutical preparationcomprising a pharmaceutically acceptable vehicle and a purifieddipeptide having the amino acid sequence L-Glu-L-Trp, or a salt of saiddipeptide, in an amount sufficient to cause said increase.
 2. The methodof claim 1 wherein the amount is 0.1 μg/kg to 1 μg/kg body mass of theanimal.
 3. The method of claim 1 wherein the amount is 1 μg/kg body massof the animal.
 4. The method of claim 1 wherein the animal suffers fromradiation induced immune deficiency.
 5. The method of claim 1 whereinthe animal suffers from staphylococcal pyoderma.
 6. The method of claim1 wherein the animal suffers from a purulent-inflammatory disease of thebone or soft tissue, thyomegalia, thymectomy, rheumatoid arthritis orfurunculosis.
 7. The method of claim 1 wherein the animal suffers froman immune deficiency condition associated with cancer.
 8. The method ofclaim 1 wherein the pharmaceutical composition is administeredintramuscularly or intranasally.
 9. The method of claim 1 wherein thepharmaceutical composition is administered intravenously.
 10. The methodof claim 1 wherein the amount of dipeptide or salt thereof is about 500μg administered in several doses.
 11. The method of claim 1 wherein thepharmaceutical preparation is administered in the form of an injectablesolution.
 12. The method of claim 1 wherein the pharmaceuticalpreparation is administered in the form of a tablet, a supporitorium, acapsule or an ointment.
 13. The method of claim 1 wherein thepharmaceutically acceptable vehicle comprises an aqueous solution ofsodium chloride, a novocain solution, distilled water, Ringer's solutionor a solution of glucose.
 14. The method of claim 1 wherein thepharmaceutically acceptable vehicle comprises starch, glucose orglycine.